Table 4. Plasmids available for transgene expression using the one-promoter system.

Plasmida	Promoterb	Upstream Genec	Selectable Markerd
pMO449e	HSP70A/RBCS2	CrVENUS-3FLAG	APHVIII
pMO470e	HSP70A/RBCS2	CrVENUS-3FLAG	APHVIII
pMO471e	HSP70A/RBCS2	CrVENUS-3FLAG	APHVIII
pMO488	HSP70A/RBCS2	CrVENUS	APHVIII
pMO490	HSP70A/RBCS2	sfGFP-3FLAG	APHVIII
pMO519	HSP70A/RBCS2	CrmCHERRY	APHVIII
pMO520	HSP70A/RBCS2	CrmCHERRY-3FLAG	APHVIII
pMO507	PSAD	CrVENUS-3FLAG	APHVIII
pMO508	TUB2	CrVENUS-3FLAG	APHVIII
pMO561f	TUB2	CrVENUS-3FLAG	APHVIII

^aAvailable plasmids with one-promoter expression constructs of the general structure shown in Figure 1B.

^bExcept as indicated in note ^f, all promoters are followed by the first three codons and intron of RBCS2.

^cAll genes indicated have been codon optimized for Chlamydomonas. These genes can be replaced in toto by HpaI–StuI fragments containing other GOIs whose expression/overexpression is desired. Fluorescence tagging of a gene product of interest can be achieved by inserting the coding sequence, in frame, at the HpaI site (C-terminal tagging) or the StuI site (N-terminal tagging).

^dOther selectable markers could be inserted as NdeI–BamHI fragments. To date, however, we have not had success with selectable markers other than APHVIII (see text).

^eThese plasmids differ only in the short linker between CrVENUS-3FLAG and APHVIII (see Figure 2D and Table 2).

^fSame as pMO508 except that the RBCS2 ATG start codon has been changed to TTG. This allowed expression of CrVENUS-3FLAG, presumably from its own start codon (i.e., without the additional amino acids from RBCS2).